This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles

for the
Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

Designation: D5338 − 15

Standard Test Method for

Determining Aerobic Biodegradation of Plastic Materials
Under Controlled Composting Conditions, Incorporating
Thermophilic Temperatures1
This standard is issued under the fixed designation D5338; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.

1. Scope* 2. Referenced Documents

1.1 This test method determines the degree and rate of
aerobic biodegradation of plastic materials on exposure to a
controlled-composting environment under laboratory
conditions, at thermophilic temperatures. This test method is
designed to yield reproducible and repeatable test results under
controlled conditions that resemble composting conditions,
where thermophilic temperatures are achieved. The test sub-
stances are exposed to an inoculum that is derived from
compost from municipal solid waste. The aerobic composting
takes place in an environment where temperature, aeration and
humidity are closely monitored and controlled.
NOTE 1—During composting, thermophilic temperatures are most
readily achieved in large-scale, professionally-managed facilities.
However, these temperatures may also be reached in smaller residential
composting units, frequently referred to as “backyard” or “home” com-
posting.
1.2 This test method is designed to yield a percentage of
conversion of carbon in the sample to carbon dioxide. The rate
of biodegradation is monitored as well.
1.3 This test method is designed to be applicable to all
plastic materials, which are intended to be composted in
facilities that achieve thermophilic temperatures.
1.4 The values stated in SI units are to be regarded as the
standard.
1.5 This standard does not purport to address all of the
safety concerns, if any, associated with its use. It is the
responsibility of the user of this standard to establish appro-
priate safety and health practices and determine the applica-
bility of regulatory limitations prior to use. Specific hazard
statements are given in Section 8.
1.6 This test method is equivalent to ISO 14855.
1
This test method is under the jurisdiction of ASTM Committee D20 on Plastics
and is the direct responsibility of Subcommittee D20.96 on Environmentally
Degradable Plastics and Biobased Products.
Current edition approved June 1, 2015. Published June 2015. Originally
approved in 1992. Last previous edition approved in 2011 as D5338 - 11. DOI:
10.1520/D5338-15.
2.1 ASTM Standards:2
D618 Practice for Conditioning Plastics for Testing
D883 Terminology Relating to Plastics
D1293 Test Methods for pH of Water
D2908 Practice for Measuring Volatile Organic Matter in
Water by Aqueous-Injection Gas Chromatography
D3590 Test Methods for Total Kjeldahl Nitrogen in Water
D4129 Test Method for Total and Organic Carbon in Water
by High Temperature Oxidation and by Coulometric
Detection
E260 Practice for Packed Column Gas Chromatography
E355 Practice for Gas Chromatography Terms and Relation-
ships
2.2 APHA—AWWA—WPCF Standards:
2540 D Total Suspended Solids Dried at 103 to 105°C3
2540 E Fixed and Volatile Solids Ignited at 550°C3
2.3 ISO Standard:
ISO 14855 Plastics—Evaluation of the Ultimate Aerobic
Biodegradability and Disintegration Under Controlled
Composting Conditions—Method by Analysis of Re-
leased Carbon Dioxide4
3. Terminology
3.1 Definitions—Definitions of terms applying to this test
method appear in Terminology D883.
4. Summary of Test Method
4.1 This test method consists of the following:
4.1.1 Selection of plastic material for the determination of
the aerobic biodegradability in a controlled-composting
system,
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
Standards volume information, refer to the standard’s Document Summary page on
the ASTM website.
3
Standard Methods for the Examination of Water and Wastewater, 17th Edition,
1989, American Public Health Association, 1740 Broadway, New York, NY 19919.
4
Available from American National Standards Institute (ANSI), 25 W. 43rd St.,
4th Floor, New York, NY 10036, http://www.ansi.org.

*A Summary of Changes section appears at the end of this standard

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 D5338 − 15
4.1.2 Obtaining an inoculum from composted municipal
solid waste,
4.1.3 Exposing the test substances to a controlled aerobic
composting process in conjunction with the inoculum,
4.1.4 Measuring carbon dioxide evolved as a function of
time, and
4.1.5 Assessing the degree of biodegradability.
4.2 The percentage of biodegradability is obtained by de-
termining the percentage of carbon in the test substance that is
converted to CO2 during the duration of the test. This percent-
age of biodegradability will not include the amount of carbon
converted from the test substance that is converted to cell
biomass and that is not, in turn, metabolized to CO2 during the
course of the test.
4.3 The disintegration of a compact test material is visually
determined at the end of the test. Additionally, the weight loss
of the test material may be determined.
5. Significance and Use
5.1 Biodegradation of a plastic within a composting unit is
an important phenomenon because it may affect the decompo-
sition of other materials enclosed by the plastic and the
resulting quality and appearance of the composted material.
Biodegradation of plastics will also allow the safe disposal of
these plastics through large, professionally-managed compost-
ing plants and well-run residential units, where thermophilic
temperatures are achieved. This procedure has been developed
to permit the determination of the rate and degree of aerobic
biodegradability of plastic products when placed in a con-
trolled composting process.
5.2 Limitations—Because there is a wide variation in the
construction and operation of composting facilities and be-
cause regulatory requirements for composting systems vary,
this procedure is not intended to simulate the environment of
any particular composting system. However, it is expected to
resemble the environment of a composting process operated
under optimum conditions where thermophilic temperatures
are achieved. More specifically, the procedure is intended to
create a standard laboratory environment that will permit a
rapid and reproducible determination of the aerobic biodegrad-
ability under controlled composting conditions.
6. Apparatus
6.1 Composting Apparatus (see Fig. 1):
6.1.1 A series of at least twelve composting vessels (one test
substance, one blank, one positive and one negative control, all
in three replicates) of 2 to 5 L of volume. For screening
purposes, depending upon the test material, a smaller volume
also may be used.
6.1.2 Water Baths, or other temperature controlling means
capable of maintaining the temperature of the composting
vessels at 58°C (62°C).
6.1.3 Pressurized-Air System, that provides CO2-free, H2O-
saturated air to each of the composting vessels at accurate
aeration rates. If using a direct measurement of CO2 (see 6.4),
then normal air may be used.
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FIG. 1 Set-Up Using Carbon Dioxide-Trapping Apparatus
6.1.4 Suitable devices for measuring oxygen and CO2
concentrations in the exhaust air of the composting vessels,
such as specific sensors or appropriate gas chromatographs.
6.2 Carbon Dioxide-Trapping Apparatus for Each Com-
posting Vessel:
6.2.1 At least three 5000-mL bottles fitted with gas sparging
and containing Ba(OH)2 carbon-dioxide scrubbing solution.
6.2.2 Flexible Tubing, nonpermeable to carbon dioxide.
6.2.3 Stoppers, equipped with gas-sampling parts.
6.3 Miscellaneous:
6.3.1 Analytical Balance, (61 mg) to weigh test specimen.
6.3.2 100-mL Burette.
6.3.3 0.05 N HCl.
6.3.4 pH Meter.
6.3.5 Suitable devices and analytical equipment for measur-
ing dry solids (at 105°C), volatile solids (at 550°C), volatile
fatty acids by aqueous-injection chromatography, total Kjel-
dahl nitrogen and carbon concentrations.
6.4 Optional—The carbon dioxide-trapping apparatus and
titration equipment can be replaced by a gas flow meter plus a
gas-chromatograph, or other apparatus equipped with suitable
detector and column(s), for measuring CO2 and O2 concentra-
tions in the exhaust air of each vessel. Take care to analyze
CO2 concentration on a sufficiently frequent basis in order to
produce a reliable cumulative CO2 production over the course
of the test (for example, every 3 to 6 h). A standard gas should
be injected to internally standardize the gas-chromatograph on
a continuous basis over the course of the test. Operate the gas
chromatograph in conformance with Practices E260 and E355
(see Fig. 2).
6.5 Ensure that all glassware is cleaned thoroughly and free
from organic matter.
7. Reagents and Materials
7.1 Barium Hydroxide Solution, approximately 0.024 N and
then standardized, prepared by dissolving 4.0 g Ba(OH)2 per
litre of distilled water. Filter through filter paper and store
sealed as a clear solution to prevent absorption of CO2 from the
air.
 D5338 − 15
FIG. 2 Optional Set-Up Using a Gas Chromatograph
7.2 Analytical-Grade Cellulose, for thin-layer chromatogra-
phy with a particle size of less than 20 µm as positive control.5
7.3 Polyethylene, as a negative control. It should be in the
same form as the form in which the sample is tested (polyeth-
ylene film for film samples, polyethylene pellets in case sample
is in the form of pellets, etc.).
8. Hazards
8.1 This test method requires the use of hazardous chemi-
cals. Avoid contact with the chemicals and follow manufactur-
er’s instructions and Material Safety Data Sheets.
8.2 The compost inoculum may contain sharp objects. Take
care when handling it.
8.3 The composting vessels are not designed to withstand
high pressures. The system should be operated at close to
ambient pressure.
9. Compost Inoculum
9.1 The compost inoculum should be two to four months old
well-aerated compost coming from the organic fraction of
municipal solid waste and sieved on a screen of <10 mm. If
such a compost is not available, compost from plants, treating
green, or yard waste, or mixtures of green waste and municipal
solid waste may be used. It is recommended that the compost
inoculum produces between 50 and 150 mg of CO2 per gram of
volatile solids over the first ten days of the test, and has an ash
content of less than 70 % and a pH between 7 and 8.2. Total dry
solids should be between 50 and 55 %.
9.2 The compost inoculum should be as free from larger
inert materials (glass, stones, metals, etc.) as possible. These
items should be removed manually as much as possible to
produce a homogeneous compost inoculum.
9.3 It is recommended to use compost of sufficient porosity
to enable conditions to be as aerobic as possible. Addition of
structural material, such as small wood particles, or persistent
or poorly biodegradable inert material may prevent the com-
post from sticking together and clogging during the test.
5
For development of this test method, Avicel, available from EM Chemicals,
Inc., Hawthorne, New York, was used.
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10. Test Specimen
10.1 The test specimen should have sufficient carbon to
yield carbon dioxide that can be adequately measured by the
trapping apparatus or CO2 measurements.
10.2 All basic composting parameters, such as C/N, oxygen
in the composting vessel, porosity, and moisture content should
be optimized so as to make a good composting process
possible. The C/N ratio should preferably be between 10 and
40 for both the inoculum and test substance combined. Oxygen
levels in the composting vessel should be at least 6 % at all
times and no free-standing water nor clumps of material should
be present.
10.3 Test specimens may be in the form of films, formed
articles, dog bones, granules, powder, or other, and conform to
Practice D618.
11. Procedure
11.1 Preparation of the Samples:
11.1.1 Obtain an inoculum from a properly operating aero-
bic composting plant treating municipal solid waste, or the
organic fraction thereof. If required, further stabilize the
inoculum at the laboratory in order to obtain a low CO2
production (see 9.1.).
11.1.1.1 Screen the inoculum to less than 10 mm and
manually remove and discard any large inert items (pieces of
glass, stone, wood, etc.). Determine volatile solids, dry solids
and nitrogen content according to Test Methods D3590,
D1888, and APHA Test Methods 2540 D and 2540 E.
11.1.2 Determine volatile solids, dry solids and carbon
content of all the test substances according to APHA Test
Methods 2540 D and 2540 E and Test Method D4129.
11.1.3 Weigh out roughly 600 g of dry solids of inoculum
and mix with about 100 g of dry solids coming from the
sample. Adjust the dry solids content of the mixture in the
vessel to approximately 50 % with distilled water. Add ammo-
nium chloride if the C/N ratio is more than 40. Weigh vessels
with all of the contents immediately before initiation of the
composting process.
11.1.4 The blank consists of the inoculum only, containing
about 600 g of dry solids. As references, use thin-layer
chromatography cellulose as a positive control and polyethyl-
ene as a negative control.
11.1.5 The test material may be in the form of films, formed
articles such as dog bones, granules, or powder. The maximum
surface area of a compact test material used should be about 2
by 2 cm. In case the original test material is larger, reduce it in
particle size.
11.1.6 No more than about 3⁄4 of the volume of the test
vessel should be filled with test mixture. Sufficient headspace
is required in order to provide enough space for manual
shaking of the test mixture.
11.2 Start-Up Procedure—Initiate aeration of the compost-
ing vessels with air-flow rates that are sufficiently high to
ensure that oxygen levels do not drop below 6 % in the exhaust
air. Oxygen levels should be closely controlled during the first
week and measured at least twice daily. Adjust air-flow rates as
needed.
 D5338 − 15
11.3 Operating Procedure:
11.3.1 The composting vessels are incubated in the dark or
in diffuse light for a period of 45 days in an enclosure that is
free from vapors toxic to microorganisms. The temperature is
maintained at 58°C (62°C). In special cases, for example,
when the melting point of the test material is low, another
temperature may be chosen. This temperature should be
constant during the test and kept in a range of 62°C. The
change of temperature should be justified and clearly indicated
in the test report.
11.3.2 Check CO2 and O2 concentrations in the outgoing air
at least daily with a minimum time interval of 6 h after the first
week for the remainder of the test.
11.3.3 Check air flow daily before the composting vessels
and at the outlets, ensuring that no leaks are present in the
complete system. Adjust air flow to maintain a CO2 concen-
tration of at least 2 % volume over volume to allow accurate
determination of CO2 level in the exhaust air.
11.3.4 Ensure proper composting conditions. Shake the
composting vessels weekly to prevent extensive channelling,
provide uniform attack on the test specimen and provide an
even distribution of moisture. In case excessive moisture levels
are observed, such as free-standing water in the vessels or
clumping due to high moisture content, remove excess liquid
by injecting dry air, or by drainage via air inlet. If excessively
dry conditions are observed, that will severely slow down the
breakdown process, add moisture. During the whole course of
the test, make adjustments to ensure proper composting con-
ditions. If adjustments are made, then CO2 and O2 concentra-
tions must be monitored closely during the following 72 h and
measured at least twice daily with a time interval of more than
6 h.
11.3.5 At the weekly shaking and at the end of the test,
record visual observations with regard to compost structure,
moisture content and color, fungal development, smell of the
exhaust air, and sample disintegration.
11.3.6 The incubation time of 45 days may be extended if
significant biodegradation of the test substance is still being
observed.
11.4 End of the Test:
11.4.1 At the end of the test, weigh the vessels with the
contents and determine the dry solids concentration remaining
in the composted material. Volatile solids may be determined if
weight loss is to be calculated.
11.4.2 Measure the pH in conformance with Test Methods
D1293. If the pH is less than 7, measure the volatile fatty acids
spectrum to indicate souring of the contents in the composting
vessel in accordance with Practice D2908. Measure the pH by
diluting the sample on a 5:1 w/w ratio of distilled water to
compost inoculum or residue, mix by shaking manually and
measure immediately.
11.4.3 If more than 2 g of volatile fatty acids per kilogram
of dry matter in the composting vessel is formed, the test must
be regarded as invalid.
12. Calculation
12.1 Determine the total carbon content of the test material
by elemental analysis or by calculation if the chemical com-
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position is well established. This allows the theoretical quantity
of carbon dioxide evolution to be calculated as follows:
material 5 w % carbon
w/100 3 g of material charged 5 Y g carbon charged to
compost vessel 5 C i
C1O 2 →CO2
12 g C yields 44 g CO2
44 3 Y
Y g C yields g CO2
12
12.2 Determine the cumulative CO2 production (in grams)
from the test substances.
12.2.1 Determine the amount of CO2 produced by the
difference, in millilitres of titrant, between the test substance
and blank Ba(OH)2 traps. Perform the titration with 0.05 N
HCl.
12.2.1.1 When CO2 enters the absorber bottles, it reacts in
the following manner:
Ba~ OH! 2 1CO2 →BaCO3 1H 2 O
12.2.1.2 The BaCO3 formed is insoluble and precipitates.
Determine the amount of Ba(OH)2 remaining in solution by
end-point titration with HCl using phenolpthalein as an indi-
cator according to the following equation:
Ba~ OH! 2 12 HCl→BaCl2 12 H 2 O
12.2.1.3 From the above two equations, it can be seen that
the number of mmol of CO2 produced is:
mmoles HCl
mmoles of CO2 5 mmoles of Ba~ OH! 2 at start 2
2
12.2.2 For the option with gas chromatography, the cumu-
lative CO2 production (in grams) is determined from the
measurements of flow rate and gas composition and after
recalculation to STP (standard temperature and pressure)
conditions.
12.2.3 Calculate the amount of cumulative gaseous-carbon
produced by each reactor.
12.2.4 Determine the mean (of the three replicates) net
gaseous-carbon production by controlled composting of the
test substances by subtracting the mean gaseous carbon pro-
duction of the control (three replicates) containing only the
inoculum.
12.3 Calculate the percent of biodegradation by dividing the
average net gaseous-carbon production of the test compound
by the original average amount of carbon in the test compound
and multiplying by 100:
mean C g ~ test! 2 mean C g ~ blank!
% biodegradation 5 3 100
Ci
where:
Cg = amount of gaseous-carbon produced, g, and
Ci = amount of carbon in test compound added, g.
12.4 Calculate the standard error, se, of the percentage of
biodegradation as follows:
2
S e 5 SQRT~~ s test/n1 ! 1 ~ s 2blank/n2 !! 3 100/C i
n1 and n2 are the number of replicate test and control di-
gesters respectively; s is the standard deviation of the total
gaseous carbon produced.
 D5338 − 15
12.5 Calculate the 95 % confidence limits as follows: 14.1.9 Results on the visual observations of the compost
95 % CL 5 % biodegradation6 ~ t 3 s e ! inoculum and the test material during and at the end of the test,
Where t is the t-distribution value for 95 % probability with such as moisture content, fungal development, structure, color,
(n1 + n2 − 2) degrees of freedom; thus n = 3 + 3 − 2 = 4. and smell,
14.1.10 Describe qualitatively the disintegration of the test
13. Interpretation material in the case of a compact test material. Add further
information, such as photographs or measured physical values,
13.1 Information on the toxicity of the plastic material may
if these are available, and
be useful in the interpretation of inhibitive effects.
14.1.11 Results on weight measurements of compost vessels
13.2 In most instances when investigating a plastic material, at the start and at the end of the test, and test material weight
a reference or control substance known to biodegrade is loss, if determined.
necessary in order to check the activity of the inoculum. If
sufficient biodegradation (a minimum of 70 % for cellulose 15. Precision and Bias
within 45 days) is not observed with the positive reference, the 15.1 The precision and bias of the procedure in this test
test must be regarded as invalid and should be repeated, using method is being determined.
new inoculum. If the deviation of the percentage of biodegra- 15.2 Preliminary results for within-laboratory repeatability
dation of the positive reference is greater than or equal to 20 % testing using a controlled composting set-up with gas chro-
at the end of the test, then the test shall be regarded as invalid. matograph are presented in Table 1. These data represent three
different determinations of the degradation of cellulose as a
14. Report positive reference. The average degradation of cellulose after
14.1 Report the following data and information: 45 days of composting at a constant temperature of 50°C
14.1.1 Information on the inoculum, including source, per- (62°C) was 75.3 %, with an average standard deviation of
cent dry solids, percent volatile solids, total Kjeldahl nitrogen, 2.5 % and an average 95 % confidence limit interval of 5 %.
activity (CO2 production in the first ten days), date of All three runs were carried out within a seven-month period by
collection, storage, and handling. the same operators. Fig. 3, 4, and 5 represent a graphical view
of the third run in which a biodegradability of 78.9 % was
14.1.2 Carbon content of the plastic materials, positive and
obtained as the mean for the three replicates containing
negative control, and calculation of the theoretical maximum
cellulose as the positive control, with a standard deviation of
carbon dioxide production. Report specific information on the
0.3 % and a 95 % confidence limit interval of 2 %. The graphs
size, shape, volume, and thickness of the plastic materials
represent the results from three replicates for the inoculum only
tested, along with the form of the plastic material, that is, sheet,
as the blanks (see Fig. 3), the cellulose as the positive control
powder, pellet, etc.,
(see Fig. 4) and the net CO2 production per gram of cellulose
14.1.3 Weight of vessels with contents before and at the end added (see Fig. 5).
of the test,
14.1.4 Cumulative carbon dioxide evolution and oxygen 16. Keywords
consumption over time and display graphically. Report appa- 16.1 aerobic biodegradation; biodegradation; composting;
ratus used for carrying out the test method, plastics
14.1.5 Percentage of aerobic biodegradation for each plastic
material tested, the standard deviation and 95 % confidence TABLE 1 Results from Within-Laboratory Testing for the Aerobic
interval for each material or control substance tested for the Biodegradability of Cellulose as a Positive Control Under
percent of biodegradation, Controlled Composting Conditions
14.1.6 Percentage of biodegradation relative to the positive Biodegradability Standard 95 % Confidence
after 45 days, % Deviation, % Limit, %
reference (cellulose = 100 %),
Run 1 76.7 4.9 8.2
14.1.7 Temperature range of test, Run 2 70.3 2.4 4.8
14.1.8 pH of compost inoculum and pH of final residues. Run 3 78.9 0.3 2.0
Mean of 75.3 2.5 5.0
Volatile fatty acids concentration for vessels with a final pH of Three Runs
less than 7,

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 D5338 − 15

FIG. 3 Cumulative CO2 Production from Inoculum

FIG. 4 Cumulative CO2 Production from Inoculum Plus Cellulose

FIG. 5 Net Cumulative CO2 Production from Cellulose

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 D5338 − 15
SUMMARY OF CHANGES

Committee D20 has identified the location of selected changes to this standard since the last issue (D5338–11)
that may impact the use of this standard. (June 1, 2015)

(1) Revised 6.3.1.

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